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Showing: 551 - 600 of 954 results

  1. CRISPR Antimicrobials

    Type
    Blog Post
    ...virulent or antibiotic resistant bacteria, and inappropriate antibiotic usage selects for resistant bacteria...to combat antibiotic resistance using multiple approaches. Major restaurant chains including Chipotle and...bacterial host range. To further potential clinical applications, Ando et al. recently engineered synthetic phages...

  2. Tips for Screening with Yeast Two Hybrid Systems

    Type
    Blog Post
    ...or growth stage). A prey library can either be applied to the bait in a single population (e.g. transforming...although these can often be accounted for using appropriate replicates and controls (see Tips below), and.... Obrdlik, P., El-Bakkoury, M., Hamacher, T., Cappellaro, C., Vilarino, C., Fleischer, C., Ellerbrok, ...

  3. Cpf1: A New Tool for CRISPR Genome Editing

    Type
    Blog Post
    ..., Zetsche et al. developed a Cpf1 multiplexing approach using a single crRNA array to express up to 4 ...-target editing in a cell culture model. The application of Cpf1 to genome editing is exciting both in...in terms of basic science and translational applications. This discovery of this type V CRISPR system ...

  4. The Importance of a Fun Workplace: Company Culture at Addgene

    Type
    Blog Post
    ... result seems to be a balance of productive and happy employees. The teambuilding culture helps us build...throughout the years. This team building merriment happened naturally when Addgene was a family affair back... sure people know that there is something fun happening! Addgene actually has chimes outside of our break...

  5. Plasmids 101: Shuttle Vectors

    Type
    Blog Post
    ...which helps increase cellular uptake, but this approach requires compatible cell lines that express viral...individual analysis. A shuttle vector designed for this approach in a mammalian cell experiment should still include...terms are typically used in different (though overlapping) ways. Broad host range plasmids are those that...

  6. Hot Plasmids Spring 2024

    Type
    Blog Post
    ...Paul Blainey’s lab has developed a multiplexing approach to Cas9-based pooled CRISPR screens called CROPseq-multi...for microglia research and broader therapeutic applications.  Find microglia-targeting AAV plasmids here...reconstituting coding sequences for a range of applications, including CRISPR activation by dCas9-VPR, prime...

  7. New Acoustic Reporter Genes: Ultrasound Imaging of Gene Expression

    Type
    Blog Post
    ... the bacteria were injected, limiting their applicability. They also did not scatter ultrasound nonlinearly...chromatic-opening agents to boost expression to appreciable levels. We wanted to create second-generation...ultrasound imaging of nanoscale acoustic biomolecules. Appl. Phys. Lett. 110, 073704 (2017). Hurt, R. C. et ...

  8. Changing Labor Laws Bring Increased Postdoc Wages

    Type
    Blog Post
    ...institutions, or institutions they're thinking about applying to, are doing - for example, your benefits may...get rid of postdocs; that current postdoctoral appointments may shorten in length; that postdoc salaries...trainee workforce has clearly not been a sustainable approach to biomedical research in the U.S. and postdoctoral...

  9. AAV Titers: Where Do They Come From And What Do They Mean?

    Type
    Blog Post
    ...used to generate AAV.  One study showed that approximately 50% of viral capsids were genome-containing ...particles can be identified after negative staining and appear darker than full vector particles. This image shows...experiments? Because (physical) AAV titers are approximations and they depend highly on the experimental ...

  10. Advanced Uses of Cre-lox and Flp-FRT - A Neuroscientist’s View

    Type
    Blog Post
    ... its reintroduction, inversion and re-inversion happen at the same probability. Continuous inversion of...the same time from two different plasmids this approach becomes problematic as it reduces the probability... Cell Types for Imaging and Long-Range Circuit Mapping.” Journal of Neuroscience 28(28): 7025–30. PubMed...

  11. Rett Syndrome: A History of Research and Therapeutic Outlooks

    Type
    Blog Post
    ...highlights some of the latest breakthrough therapeutic approaches. What is Rett Syndrome? Caused by a mutation ...technical challenges to overcome for both of these approaches, such as delivery of the editing machinery to...Extends the Survival of MeCP2-Null Mice without Apparent Toxicity after Intracisternal Delivery. Molecular...

  12. Deciphering the Mysteries of Behavior with Viral Vectors

    Type
    Blog Post
    ...movement in an open field box or a circular corridor apparatus using a camera or tracking software. Locomotion...and might open new ways for a novel therapeutic approach against drug addiction. Anxiety tests Rodents...measuring the time spent in a particular part of the apparatus, it turned out that the IL-1 beta overexpressing...

  13. The Challenges of Cell Culture

    Type
    Blog Post
    ...especially with industry. Today we also have the appropriate tools to verify our cells lines. Even if you ...the results are reliable. With this two-pronged approach, the many mistakes of the past can be prevented...including GSK, AstraZeneca-MedImmune, and Infors & Applikon. Due to his high level of experience he now travels...

  14. New Optogenetic Tools for Cytoskeleton and Membrane Control

    Type
    Blog Post
    ... barrier of the endothelium, but they could be applied to study other actin-dependent processes in cells...) and to introduce unnatural head groups for applications like fluorescent labeling via click chemistry...constructs are great for in vitro biocatalytic applications where toxicity doesn’t matter. Plus, for folks...

  15. CRISPR 101: Validating Your Genome Edit

    Type
    Blog Post
    ... explain how to verify that your cells were appropriately edited for your desired mutation - insertion...frequency is 50%, then you could infer that approximately 25% of cells are likely -/-, 50% are -/+, and...sequencing confirmation. All knock-ins A method applicable to knock-ins both large and small is a software...

  16. Fluorescent Proteins 101: GFP Fusion Proteins - Making the Right Connection

    Type
    Blog Post
    ...study its function is one of the most popular applications of fluorescent proteins. These fusion proteins...targeted to the endoplasmid reticulum based on the appearance of OSER whorls. This assay is quantitative and...linker sequence short In the early days of GFP applications, many were concerned with steric hindrance by...

  17. The Fluorescent Vegetables in Aptamer Soup

    Type
    Blog Post
    ...enjoy a warm meal on a winter day, you should be happy to learn that many DNA-based reagents are named ...aptamer soup by exploring the development and applications of fluorescent RNA aptamers in Addgene’s collection...fluorescence when bound to DFHBI despite being approximately half the length (Warner et al., 2014); its length...

  18. Harnessing Bacterial Toxins for Allelic Exchange

    Type
    Blog Post
    ...References Espah Borujeni, Amin, Anirudh S. Channarasappa, and Howard M. Salis. "Translation rate is controlled...Scarless Modification of Proteobacterial Genomes." Applied and environmental microbiology (2019): AEM-00990...chromosomal gene replacement in Escherichia coli." Appl. Environ. Microbiol. 81.15 (2015): 5103-5114. PubMed...

  19. Multiple Plasmids at a Low Price: Kits from Addgene

    Type
    Blog Post
    ...polycistronic E. coli expression constructs. Some applications of plasmids from this kit include the production...can be engineered to bind specific RNA bases. By swapping out these different modular repeat segments, you...variety of functions via different fusions. Some applications of PUF protein fusions reported in the literature...

  20. Pouring LB Agar Plates

    Type
    Protocol
    ...concentration disolved in the appropriate liquid solvent. See table to the right for appropriate antibiotic concentrations...Strains Grow Assuming the appropriate strains were streaked on the appropriate plates, then if both strains... LB-agar powder you’ve measured out into an appropriately sized bottle for autoclaving. We make 400 mL...autoclave. Be sure to check the literature for an appropriate sterilization technique if you are working with... wipe down with a paper towel. Count out the appropriate number of plates and stack them on your lab bench...of molten gel mix, a tube rack containing the appropriate antibiotics, and a section for active pouring...antibiotic. Incubate both plates overnight at the appropriate growth temperature and check for growth. See ...

  21. Hot Plasmids: Spring 2025

    Type
    Blog Post
    ...popular choice for a variety of synaptic imaging applications. Find iGluSnFR4 plasmids here! Aggarwal, A....that they could reprogram the genomic target by swapping the homology arms for new targets. And, importantly... is scarless, unlike previous integrase-based approaches. Find STITCHR plasmids here! Fell, C. W., Villiger...

  22. Viral Vectors 101: Optogenetic Tools

    Type
    Blog Post
    ...tools Once you’ve selected the class of tool appropriate for your experiment, you’ll need to find a specific...Yizhar, O., & Deisseroth, K. (2012). Principles for applying optogenetic tools derived from direct comparative...

  23. Plasmids 101: Repressible Promoters

    Type
    Blog Post
    ...for sphingomonads and other Alphaproteobacteria. Appl Environ Microbiol. 2013 Nov;79(21):6795-802. doi...cumate-inducible gene expression system for Bacillus. Appl Microbiol Biotechnol. 2019 Jan;103(1):303-313. doi...expression of genes for metabolic engineering applications in Saccharomyces cerevisiae.” FEMS Yeast Res...

  24. Technologies Enabled by NanoLuc® Luciferase

    Type
    Blog Post
    ...et al (2). In this post, I’ll cover two great applications of the NLuc technology. The first couples NLuc...the coupled luciferase. This strategy was first applied using RLuc8, an engineered Renilla luciferase, ...fluorescence. The paper reports a wide variety of applications of the Enhanced Nano-Lanterns including monitoring...

  25. CRISPR 101: Epigenetics and Editing the Epigenome

    Type
    Blog Post
    ...to specific DNA sequences. Non-editing CRISPR applications direct catalytically dead dCas9 fused to a variety... publication at Addgene. Klann TS, Black JB, Chellappan M, Safi A, Song L, Hilton IB, Crawford GE, Reddy...Ruan K, Wang F, Xu G, Hu R (2016) A CRISPR-based approach for targeted DNA demethylation. Cell Discovery...

  26. Quick Guide to All Things Lentivirus

    Type
    Blog Post
    ...have been increasingly popular in fundamental and applied research since their first use in the early 90’... at transferring the gene of interest to the appropriate cells, it was obvious that its safety was a disadvantage...either tTA or rtTA (FUW-M2rtTA). In this kind of approach, a population of cells might express only the ...

  27. Transferable Skills Guide: Teamwork

    Type
    Blog Post
    ...cross-team guy” at Addgene, an unofficial title that I happily embrace (and admittedly, I may have started that...identify as one.  But some time around 10th grade, I snapped out of the phase of my life that had me listening...you’re part of a team - it will just make you a happier person. What does a good team need? Besides that...

  28. Protocol - Bacterial Transformation

    Type
    Protocol
    ... thaw on ice (approximately 20-30 mins). Remove agar plates (containing the appropriate antibiotic ) from...spreading device Reagents LB agar plate (with appropriate antibiotic) LB or SOC media Competent cells DNA...Pro-Tip Transformation efficiencies will be approximately 10-fold lower for ligation of inserts to vectors...transformation onto a 10 cm LB agar plate containing the appropriate antibiotic. Pro-Tips We recommend that you plate... take up the DNA, an electro-magnetic field is applied to the cell/DNA mixture to induce membrane permeability... to have access to an electroporator and the appropriate cuvettes. Follow the manufacturer's instructions...

  29. Protocol - pLKO.1 – TRC Cloning Vector

    Type
    Protocol
    ...your convenience. See “warranty information” in appendix. Table of Contents A. pLKO.1-TRC Cloning Vector...References H.1 Published articles H.2 Web resources I. Appendix I.1 Sequence of pLKO.1 TRC-Cloning Vector I.2 ...GenHunter: #Q401 FuGENE® 6 Transfection Reagent Roche Applied Biosciences: #11814443001 OPTI-MEM® serum-free ...protamine sulfate, and puromycin are located in the “Appendix”. F.2. Determining the Optimal Puromycin Concentration...overnight. Day 2: b. The target cells should be approximately 80-90% confluent. c. Dilute puromycin in the...increments. d. Label plates from 1-10 and add appropriate puromycin-containing media to cells. Days 3+:...overnight. Day 2: b. Target cells should be approximately 70% confluent. Change to fresh culture media...

  30. CRISPR 101: Cytosine Transversion Editors

    Type
    Blog Post
    ...huge innovation, there was a limitation to this approach. CBEs and ABEs are base transition editors. They...GBEs), Xueli Zhang’s lab, returned with a unique approach to improving C → G editing. Instead of fusing ...abasic site. In this post, it refers to the same approach as a CGBE. BER Base Excision Repair A cellular...

  32. CRISPR 101: Multiplex Expression of gRNAs

    Type
    Blog Post
    ...chance that all the edits you want to make will happen.  There are many reasons to multiplex gRNAs, some...adjacent to the sites. When digested with the appropriate Type IIS enzyme, the unique flanking overhang...subsequent “bricks” to be added one by one. This approach is especially useful for combinatorial analyses...

  33. New Antibodies in the Lab Animation!

    Type
    Blog Post
    ...Thinking about using antibody-based applications in your experiment...but not sure where to start or ...or quick refresher on — these common antibody applications.  Thanks for watching, and we hope you found...

  34. #DeckTheLab to Win a Tasty Winter Treat This Holiday Season

    Type
    Blog Post
    ...wintertime festivities! Help us spread some cheer by snapping a photo of you and your labmates celebrating the... the New Year. Will your lab be a lucky winner? Happy holidays from all of us at Addgene! We look forward...

  35. 'Tis the Season to #DeckTheLab

    Type
    Blog Post
    ... Why #DecktheLab? Help us spread some cheer by snapping a photo of you and your labmates celebrating the... the New Year. Will your lab be a lucky winner? Happy holidays from all of us at Addgene! We look forward...

  36. Isolating a Monoclonal Cell Population by Limiting Dilution

    Type
    Protocol
    ...pool has been sufficiently selected with the appropriate antibiotic so that every cell in the culture ...stable cells such that the next day they will be approximately 50–60% confluent. For 293T cells this is about...confluent or the medium is acidified (i.e., the medium appears orange/yellow if using a standard phenol red pH...counting the cells. When you ultimately transfer approximately 50–100 cells to the conditioned medium, it will...solution at a concentration of 5 cells/mL. Prepare approximately 10 mL of this cell solution for each 96-well...which you see growth. At this time, the cells will appear as colonies in the well ( Figure 1 ). You will ...

  37. SciComm with the Experts at Science in the News Part 1

    Type
    Blog Post
    ... when Communicating with Non-Scientists How to Approach Different Audiences Listen to the SITN Podcast...Addgene Blog Check Out our Interview with Science Rapper Tom McFadden Learn about SITN's work at DayCon ...

  38. Protocol - How to Perform a Diagnostic Digest

    Type
    Protocol
    ...contains the expected size insert. By selecting the appropriate enzyme(s), one can either linearize a plasmid...interest (see below for recommended amounts) Appropriate restriction enzyme (see manufacturer's instructions...instructions for proper amount) Approrpriate restriction digest buffer (see manufacturer's instructions) Gel...others. When choosing restriction enzymes for this approach, it is often a good idea to choose two different...products on a gel and those clones with the appropriately sized products should have the correct orientation...

  39. Centrifugation

    Type
    Protocol
    ...container Personal protective equipment (PPE) appropriate for the biosafety level of the lab space Reagents... like the one in the picture above. Wear PPE appropriate for the lab space in which you are working. Even...that the centrifuge is clean and that everything appears to be working smoothly. Place your sample tubes... the centrifuge so that the pellets form in approximately the same place in each sample. Place the centrifuge...according to speed and time (and temperature, if applicable) listed in your protocol. Pro-Tip Spin speed ...

  40. Twelve Amazing Guest Blog Contributors!

    Type
    Blog Post
    ...Performing In Vivo CRISPR Screens Using the FITS Approach Caitlin Faulds, who introduced the very first...Modular Cloning and Plasmids 101: Modular Cloning Applications and Kits We are very grateful for each and...

  41. Antibody Data Repositories and Search Engines

    Type
    Blog Post
    ... specificities, from target-specific ones to application-specific ones to anything goes. We've shared ...repositories and search engines Website Targets Application Purpose Notes Antibodypedia Any Any...

  42. Deck the Lab 2022!

    Type
    Blog Post
    ...reveal the winning entries on Twitter and Instagram! Happy decorating, all, and to all a good light!    🤩 ...set a challenge to my lab mates. They did not disappoint. #deckthelab pic.twitter.com/X4eKUGnQMe— Alex...

  43. Call for Guest Bloggers

    Type
    Blog Post
    ... Molecular biologist with Biogen interested in applying CRISPR for mouse genome editing. Post: Generating...Generating Mouse Models Using CRISPR/Cas9   Erik L. Snapp, Associate Professor at Albert Einstein College of...

  45. CRISPR Cheat Sheet

    Type
    Blog Post
    ...an organism’s genome usually for a research or applied goal. Components of CRISPR: gRNAs: Sequences ...produce either a gRNA, a nuclease, or both. Applications of CRISPR: Cut: CRISPR can be used to cut the...

  47. Summer Fun at Addgene!

    Type
    Blog Post
    ... Dives” series comes in! We’ll take a common application or concept and go beyond the basics - diving ...you’re ready for the next steps on some common applications, join us as we get into the weeds on some of...

  48. AAV Packaged on Request is Here!

    Type
    Blog Post
    ... in 100 µL aliquots, so the volume options will appear as 10, 25, or 50 units.  Once your request is received... you find this service useful in your research. Happy vectoring! ...

  50. Grad School Advice Part 1: Picking a Lab and a Project

    Type
    Blog Post
    ... certain mystique that makes them difficult to approach for the youngster just starting in the lab, but... a graduate student? 4:50 - 8:00 : How did you approach doing rotations? 8:01 - 11:25 : How did you decide...
Showing: 551 - 600 of 954 results